Abstract
In an effort to identify a microbial enzyme that can be useful as a fungicide and biodegradation agent of chitinous wastes, a chitinase (Chi242) was purified from the culture supernatant of Streptomyces anulatus CS242 utilizing powder of shrimp shell wastes as a sole carbon source. It was purified employing ammonium sulfate precipitation and gel permeation chromatography techniques. The molecular weight of the purified chitinase was ~38 kDa by SDS-PAGE. The N-terminal amino acid sequence (A-P-G-A-P-G-T-G-A-L) showed close similarity to those of other Streptomyes chitinases. The purified enzyme displayed optimal activity at pH 6.0 and 50 °C respectively. It showed substantial thermal stability for 2 h at 30–60 °C, and exhibited broad pH stability in the range 5.0–13.0 for 48 h at 4 °C. Scanning electron microscopy confirmed the ability of this enzyme to adsorb onto solid shrimp bio-waste and to degrade chitin microfibers. Chi242 could proficiently convert colloidal chitin to N-acetyl glucosamine (GlcNAc) and N-acetyl chitobiose (GlcNAc)2 signifying that this enzyme is suitable for bioconversion of chitin waste. In addition, it exerted an effective antifungal activity towards fungal pathogen signifying its role as a biocontrol agent. Thus, a single microbial cell of Streptomyces anulatus CS242 justified its dual role.
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References
Ai H, Wang F, Xia Y, Chen X, Lei C (2012) Antioxidant, antifungal and antiviral activities of chitosan from the larvae of housefly, Musca domestica L. Food Chem 132:493–498
Benhabiles MS, Salah R, Lounici H, Drouiche N, Goosen MFA, Mameri N (2012) Antibacterial activity of chitin, chitosan and its oligomers prepared from shrimp shell waste. Food Hydrocoll 29:48–56
Bhatnagar A, Sillanpää M (2009) Applications of chitin- and chitosan-derivatives for the detoxification of water and wastewater—a short review. Adv Colloid Interface Sci 152:26–38
Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254
Chung Y, Tsai C, Li C (2006) Preparation and characterization of water-soluble chitosan produced by Maillard reaction. Fish Sci 72:1096–1103
Cody RM (1989) Distribution of chitinase and chitobiase in Bacillus. Curr Microbiol 19:201–205
Dahiya N, Tewari R, Hoondal GS (2006) Biotechnological aspects of chitinolytic enzymes: a review. Appl Microbiol Biotechnol 71:773–782
El-Sayed ES, Ezzat SM, Ghaly MF, Mansour M (2000) Purification and characterization of two chitinases from Streptomyces albovinaceus S-22. World J Microbiol Biotechnol 16:87–89
Green AT, Healy MG, Healy A (2005) Production of chitinolytic enzymes by Serratia marcescens QMB1466 using various chitinous substrates. J Chem Technol Biotechnol 80:28–34
Han Y, Yang B, Zhang F, Miao X, Li Z (2009) Characterization of antifungal chitinase from marine Streptomyces sp. DA11 associated with south china sea sponge Craniella Australiensis. Mar Biotechnol 11:132–140
Jayakumar R, Menon D, Manzoor K, Nair SV, Tamura H (2010) Biomedical applications of chitin and chitosan based nanomaterials—a short review. Carbohydr Polym 82:227–232
Kavitha A, Vijayalakshmi M (2011) Partial purification and antifungal profile of chitinase produced by Streptomyces tendae TK-VL_333. Ann Microbiol 61:597–603
Kim KJ, Yang YJ, Kim JG (2003) Purification and characterization of chitinase from Streptomyces sp. M-20. J Biochem Mol Biol 36:185–189
Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685
Lu S, Song X, Cao D, Chen Y, Yao K (2004) Preparation of water-soluble chitosan. J Appl Polym Sci 91:3497–3503
Miller GL (1959) Use of dinitrosalicylic acid reagent for determination of reducing sugar. Anal Chem 31:426–428
Mourya VK, Inamdar NN, Choudhari YM (2011) Chitooligosaccharides: synthesis, characterization and applications. Polym Sci Ser A 53:583–612
Mukherjee G, Sen SK (2006) Purification, characterization, and antifungal activity of chitinase from Streptomyces venezuelae P10. Curr Microbiol 53:265–269
Patil NS, Waghmare SR, Jadhav JP (2013) Purification and characterization of an extracellular antifungal chitinase from Penicillium ochrochloron MTCC 517 and its application in protoplast formation. Process Biochem 48:176–183
Pradeep GC, Yoo H, Cho S, Choi YH, Yoo JC (2015) An extracellular chitinase from Streptomyces sp. CS147 releases N-acetyl-d-glucosamine (GlcNAc) as principal product. Appl Biochem Biotechnol 175:372–386
Rabeeth M, Anitha A, Srikanth G (2011) Purification of an antifungal endochitinase from a potential biocontrol agent Streptomyces griseus. Pak J Biol Sci 14:788–797
Rinaudo M (2006) Chitin and chitosan: properties and applications. Prog Polym Sci 31:603–632
Roy I, Sardar M, Gupta MN (2003) Hydrolysis of chitin by Pectinex™. Enzyme Microb Technol 32:582–588
Shahidi F, Arachchi JKV, Jeon YJ (1999) Food applications of chitin and chitosans. Trends Food Sci Technol 10:37–51
Suresh PV (2012) Biodegradation of shrimp processing bio-waste and concomitant production of chitinase enzyme and N-acetyl-d-glucosamine by marine bacteria: production and process optimization. World J Microbiol Biotechnol 28:2945–2962
Tanabe T, Kawase T, Watanabe T, Uchida Y, Mitsutomi M (2000) Purification and characterization of a 49-kDa chitinase from Streptomyces griseus HUT 6037. J Biosci Bioeng 89:27–32
Trudel J, Asselin A (1989) Detection of chitinase activity after polyacrylamide gel electrophoresis. Anal Biochem 178:362–366
Waldeck J, Daum G, Bisping B, Meinhardt F (2006) Isolation and molecular characterization of chitinase-deficient Bacillus licheniformis strains capable of deproteinization of shrimp shell waste to obtain highly viscous chitin. Appl Environ Microbiol 72:7879–7885
Wang SL, Lin TY, Yen YH, Liao HF, Chen YJ (2006) Bioconversion of shellfish chitin wastes for the production of Bacillus subtilis W-118 chitinase. Carbohydr Res 341:2507–2515
Xiayun J, Chen D, Shenle H, Wang W, Chen S, Zou S (2012) Identification, characterization and functional analysis of a GH-18 chitinase from Streptomyces roseolus. Carbohydr Polym 87:2409–2415
Yano S, Rattanakit N, Honda A, Noda Y, Wakayama M, Plikomol A, Tachiki T (2008) Purification and characterization of chitinase A of Streptomyces cyaneus SP-27: an enzyme participates in protoplast formation from Schizophyllum commune mycelia. Biosci Biotechnol Biochem 72:54–61
Zhang J, Kopparapu NK, Yan Q, Yang S, Jiang Z (2013) Purification and characterisation of a novel chitinase from persimmon (Diospyros kaki) with antifungal activity. Food Chem 138:1225–1232
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This work was supported by research fund from Chosun University, 2015.
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Poonam Mander and Seung Sik Cho have contributed equally to this work.
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Mander, P., Cho, S.S., Choi, Y.H. et al. Purification and characterization of chitinase showing antifungal and biodegradation properties obtained from Streptomyces anulatus CS242. Arch. Pharm. Res. 39, 878–886 (2016). https://doi.org/10.1007/s12272-016-0747-3
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DOI: https://doi.org/10.1007/s12272-016-0747-3